PCR-Based Site-Specific Mutagenesis

Abstract

The alteration of gene structure through the substitution of specific nucleotides by site-specific mutagenesis is an important tool in modern recombinant DNA technology. Nucleotide changes are necessary not only for the analysis of the structural basis of gene and corresponding protein function, but also for the generation of novel gene products. The availability of the polymerase chain reaction (PCR) in the last decade has enabled the modification of DNA for different needs to be made more rapidly and easily than was previously possible. In the course of mutagenesis the relevant sequence changes can be introduced more readily by chemically synthesized oligonucleotide primers than by manipulating DNA fragments with restriction and ligation enzymes.